Hunan yunbang IPTG purification technology to achieve a breakthrough

Development background of IPTG purification:
IPTG purification process of hunan yunban achieved a breakthrough in isopropyl-catao-galactoside (IPTG, CAS No:367-93-1) is an extremely superior inducer, which is not easily metabolized by bacteria and has good stability in inducing protein expression. It is widely used in laboratory and industrial production.Because of its extensive demand, its synthesis has been studied a lot.

As a molecular biological reagent, the performance of IPTG is closely related to the product quality.Unnecessary, will seriously affect the ability to induce the existence of impurities and by IPTG synthesis process is easy to produce such as isopropyl - alpha glucosinolates isomer impurity such as galactose glucoside, and these impurities and IPTG structure and physical and chemical properties are similar, it is difficult to by conventional separation methods to eliminate, caused by the method of synthesis, or for sale on the market of IPTG inducing expression ability is weakened.Therefore, a convenient IPTG purification method is urgently needed to improve the purity and induction ability of IPTG synthesized by current methods or available in the market.

In order to solve the above technical problems, hunan yunbang invention provides a purification method of IPTG, so as to achieve the purpose of rapid separation and purification of IPTG, and improve the purity and induction ability of IPTG.

In order to achieve the above goals, hunan yunbang improved purification technology scheme is as follows:
First, IPTG and two non-soluble solvents were heated or at room temperature to make a solution with no insoluble solid in both phases. Then, the two-phase system was cooled down and IPTG crystals were precipitated.One of the two non-soluble solvents is water and the other is an organic solvent that is insoluble with water.

In the above scheme, IPTG, water and organic solvent were heated together to prepare solutions with no insoluble solids in both phases, or IPTG and water were heated for a period of time before organic solvent was added, or IPTG and organic solvent were heated for a period of time before water was added.

Scheme described in organic solvents including ethyl ether, tert-butyl methyl ether, methylene chloride, chloroform, 1, 2 - dichloroethane, methyl acetate, ethyl acetate, propyl acetate, butyl acetate, n-butyl alcohol, isobutyl alcohol, sec-butyl alcohol, chloroform, containing 4-10 carbon atoms of alkanes, petroleum ether, benzene, benzene substituents and one or several of the butanone.

The alkane containing 4-10 carbon atoms in the scheme has the following structural characteristics: CxHy, where x = 4 ~ 10, y = 8 ~ 22;Benzene containing substituents has the following structural characteristics: R is one of F, Cl, Br, I, NO2, OR1, NR2R3 and R4.R1 is a hydrogen atom, or an alkyl containing 1 ~ 10 carbon atoms;R2 and R3 are hydrogen atoms, or alkyl groups containing 1 to 10 carbon atoms;R4 is an alkyl group containing 1 to 10 carbon atoms.

The experimental heating temperature was 40 ~ 110℃, and the total mass ratio of the two non-soluble solvents to IPTG was 0.5 ~ 50:1.

Hunan yunbang IPTG purification technology has a significant impact on the industry:
Through the above technical scheme, the purification method of IPTG provided by hunan yunbang is purified by recrystallization. Crystals of different crystal types and purity can be obtained by controlling crystallization parameters.The initial nucleation has an important influence on the crystal shape and purity.

Hunan yunbang invention provides recrystallization to improve the quality of IPTG, easy to operate, easy to industrialization.In addition, the method provided by the invention is adopted to recrystallize IPTG, and the obtained IPTG products have better appearance and purity than those obtained by recrystallization of IPTG with single-phase solvent system.The obtained crystals can be used for the induction of protein expression in bacteria, and have stronger induction ability, so they have higher added value.

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